Tbx18,Wt1,and Tcf21 have been identified as epicardial markers during the early embryonic stage.However,the gene markers of mature epicardial cells remain unclear.Single-cell transcriptomic analysis was performed with the Seurat,Monocle,and CellphoneDB packages in R software with standard pro-cedures.Spatial transcriptomics was performed on chilled Visium Tissue Optimization Slides(10x Genomics)and Visium Spatial Gene Expression Slides(10x Genomics).Spatial transcriptomics analysis was performed with Space Ranger software and R software.Immunofluorescence,whole-mount RNA in situ hybridization and X-gal staining were performed to validate the analysis results.Spatial transcriptomics analysis revealed distinct transcriptional profiles and functions between epicardial tissue and non-epicardial tissue.Several gene markers specific to postnatal epicardial tissue were identified,including Msln,C3,Efemp1,and Upk3b.Single-cell transcriptomic analysis revealed that cardiac cells from wildtype mouse hearts(from embryonic day 9.5 to postnatal day 9)could be categorized into six major cell types,which included epicardial cells.Throughout epicardial development,Wt1,Tbx18,and Upk3b were consistently expressed,whereas genes including Msln,C3,and Efemp1 exhibited increased expression during the mature stages of development.Pseudotime analysis further revealed two epicardial cell fates during maturation.Moreover,Upk3b,Msln,Efemp1,and C3 positive epicardial cells were enriched in extracellular matrix signaling.Our results suggested Upk3b,Efemp1,Msln,C3,and other genes were mature epicardium markers.Extracellular matrix signaling was found to play a critical role in the mature epicardium,thus suggesting potential therapeutic targets for heart regeneration in future clinical practice.

Benign occupations of the liver (BOL) are a heterogeneous group of benign hepatic lesions,a minority of which may need active management.It's reported that radiofrequency ablation (RFA) can be used to treat BOL,such as hepatic haemangioma,focal nodular hyperplasia (FNH),hepatocellular adenoma,simple hepatic cyst,liver abscess and hepatic echinococcosis.RFA has exhibited a potentiality to replace resection as the first-line treatment option for hepatic haemangioma.For multiloculated intractable simple hepatic cyst or liver abscess,when patients are not suitable for or refuse surgical resection,RFA may be an alternative option for the resection;and it also has many advantages over resection,including effectiveness,safety,minimal invasiveness and repeatability in the treatment of FNH,hepatocellular adenoma,and hepatic echinococcosis.A review will be presented on the current situation and latest progression of RFA therapy for BOL in this article.